SNAP/CLIP-tag-Trap Agarose
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I: Input, FT: Flow-through, B: Bound
Description
SNAP/CLIP-tag®-Trap Agarose is an affinity reagent for immunoprecipitation of SNAP/CLIP-tag-fusion proteins.
It consists of a SNAP/CLIP-tag Nanobody/ VHH, which is coupled to agarose beads.
Specificity
SNAP-tag and CLIP-tag
Applications
Immunoprecipitation/ Co-IP
Mass spectrometry
On-bead enzyme assays
ChIP/ RIP analysis
| Product | Size | Code | Price | Buy |
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| Product SNAP/CLIP-tag®-Trap Agarose | Size 250 µL (10 reactions) | Code wta-10 | Price $ 275 |
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| Product SNAP/CLIP-tag®-Trap Agarose | Size 500 µL (20 reactions) | Code wta-20 | Price $ 475 |
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| Product SNAP/CLIP-tag®-Trap Agarose | Size 2.5 mL (100 reactions) | Code wta-100 | Price $ 2050 |
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| Product SNAP/CLIP-tag®-Trap Agarose | Size 5 mL (200 reactions) | Code wta-200 | Price Please inquire | |
| Product SNAP/CLIP-tag®-Trap Agarose | Size 10 mL (400 reactions) | Code wta-400 | Price Please inquire | |
| Product SNAP/CLIP-tag®-Trap Agarose Kit | Size 500 µL (20 reactions) | Code wtak-20 | Price $ 595 |
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| Product Binding Control Agarose Beads | Size 500 µL (20 reactions) | Code bab-20 | Price $ 75 |
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| Product Spin Columns | Size 10 units | Code sct-10 | Price $ 35 |
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| Product Spin Columns | Size 20 units | Code sct-20 | Price $ 60 |
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| Product Spin Columns | Size 50 units | Code sct-50 | Price $ 130 |
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| Product SNAP/CLIP-tag® VHH, recombinant binding protein | Size 250 µL | Code wt-250 | Price Please inquire |
Affinity
Dissociation constant KD of 1 nM
Wash buffer compatibility
4 M urea, 2 M NaCl, 10 mM DTT, 2 % Nonidet P40 Substitute, 1 % Triton X-100
Specificity
SNAP-tag and CLIP-tag
Coupled Nanobody/ VHH
Recombinant, monoclonal anti-SNAP/ CLIP-tag single domain antibody (sdAb) fragment
Bead properties
Bead size: ~ 90 µm (cross-linked 4% agarose beads)
Storage buffer: 20 % EtOH
Elution
- SDS sample buffer
- 0.1 mM citrate acid pH 3.0
Instead of elution, we recommend on-bead assays like on-bead digestion for MS analysis.
Compatibility with mass spectrometry
The SNAP/CLIP-tag-Trap is optimized for on-bead digestion. Complete tryptic digest results in 5-6 peptides.
Storage instructions
Shipped at ambient temperature. Upon receipt store at 4°C. Stable for one year.
Should I use an N-terminal or C-terminal fusion tag?
Both the N-terminal or C-terminal fusion tag work well with the traps.
How can I avoid unspecific protein interactions binding to the trap?
For preclearing of your sample we recommend to use our binding controls (bab-20 or bmab-20).
Please find more information in our Troubleshooting guide
How can I elute bound proteins from a trap in their native state?
You can elute your fusion protein of interest with 0.2 M glycine pH 2.5 at room temperature. Pipette the beads up and down for 60-120 seconds and repeat this step. Ensure to neutralize your supernatant immediately afterwards by adding 1 M Tris base pH 10.4.
How many mammalian cells are required for an immunoprecipitation reaction?
For one immunoprecipitation reaction, we recommend using ~10^6 - 10^7 mammalian cells. The yield is also dependent on the expression level of your protein of interest and the interaction partners.
How much cell extract should I use for an immunoprecipitation reaction?
For other type of cells than mammalian cells, we recommend using 0.5 - 1.0 mg of cell extract.
Do I need to elute bound proteins from the beads for mass spectrometry analysis?
No, you can directly conduct an on-bead digestion after immunoprecipitation. This procedure allows faster and more efficient sample preparation and a potential higher yield.
Please find more information here:
Do I need to elute my protein of interest from the beads for enzymatic assays?
No, you can directly perform your enzymatic assay on the beads if the active center is not blocked.
Please find more information in our application note "enzymatic activity assay"
What is the amount of trap slurry I need for one immunoprecipitation reaction?
25 µL slurry are sufficient for one pull-down reaction as the affinity of the traps is very high.
Benefits
- SNAP/CLIP-tag-Trap immunoprecipitates SNAP-tag- and Clip-tag- fusion proteinseven when already covalently bound to ligands such as dyes, biotin etc.
- Bound SNAP- or CLIP-tag fusion proteins can be eluted without protease
- SNAP/CLIP-tag-Trap can be used for affinity purification
- Structure and function are characterized
SNAP/CLIP-tag-Trap Agarose Kit
The SNAP/CLIP-tag-Trap Agarose is also available in a kit, including:
- SNAP/CLIP-tag-Trap Agarose
- Lysis*, wash and elution buffers
*The lysis buffer provided is suitable for mammalian cells. For other cell types use another suitable lysis buffer.
Spin Columns
- Easy and convenient handling
- Rapid washing and clean elution of bound proteins
- Simplify the pulldown of your protein
- Spin Columns for SNAP/CLIP-tag-Trap Agarose
Binding Control for Agarose Beads
- Control for unspecific binding of proteins, DNA, etc. to beads
- Pre-clearing of cell lysate
- Binding Control for SNAP/CLIP-tag-Trap Agarose
Only for research applications, not for diagnostic or therapeutic use!